American Journal of Applied Chemistry 2020; 8(1): 6-12 http://www.sciencepublishinggroup.com/j/ajac doi: 10.11648/j.ajac.20200801.12 ISSN: 2330-8753 (Print); ISSN: 2330-8745 (Online) Research of Bioactive Chemical Markers in Trunk Bark Extracts of Anogeissus leiocarpa , a Traditional Herbal Remedy for Hypertension Benjamin Ouedraogo 1, 2, Jules Yoda 1, Felix Kini 1, 2, *, Jean Claude Ouedraogo 2, Marius Lompo 1, 1 Sylvin Ouedraogo 1Department of Medicine, Traditional Pharmacopeia and Pharmacy (Mephatra/Ph), Research Institute for Health Sciences (IRSS), Ouagadougou, Burkina Faso 2Laboratory of Environmental Analytical Chemistry and Bioorganic (Lcaebio), Joseph Ki-zerbo University, Ouagadougou, Burkina Faso Email address: *Corresponding author To cite this article: Benjamin Ouedraogo, Jules Yoda, Felix Kini, Jean Claude Ouedraogo, Marius Lompo, Sylvin Ouedraogo. Research of Bioactive Chemical Markers in Trunk Bark Extracts of Anogeissus leiocarpa , a Traditional Herbal Remedy for Hypertension. American Journal of Applied Chemistry . Vol. 8, No. 1, 2020, pp. 6-12. doi: 10.11648/j.ajac.20200801.12 Received : January 21, 2020; Accepted : February 10, 2020; Published : February 18, 2020 Abstract: High blood pressure or hypertension is a public health problem in Burkina Faso. According to official statistics, its prevalence rate is estimated at 20%. The management of this disease is long-term with chronic treatments. This leads the population to resort to efficient, accessible and cheaper herbal medicines. Anogeissus leiocarpa is one of the plants used in traditional medicine for the treatment of hypertension in Burkina Faso . Earlier preclinical studies have proved the efficacy and safety of trunk bark extract. In this investigation, bioactive chemical marker phytocompounds are sought for the development, quality control and standardization of the antihypertensive drug based on this plant. The extracts of this plant are prepared by decoction from trunk bark powder and then fractionated successively with solvents in order of increasing polarity (n-hexane, dichloromethane, ethyl acetate and 1-butanol). The fractions obtained are subjected to phytochemical screening. The antioxidant properties of the different extracts have also been studied since the chemical species known as free radicals, which cause oxidative stress, are related to hypertension. The evaluation of the contents of total phenolic compounds and total flavonoids, which would have antihypertensive properties, was carried out according to the method using Folin-Ciocalteu. The highest total phenolic and total flavonoid contents were obtained with the ethyl acetate fraction (89.52 mg tannic acid equivalent/g dry extract and 70.87 mg quercetin equivalent/g dry extract for flavonoids). The best antioxidant power is also obtained with the ethyl acetate fraction (IC50: 6.93µg /mL). Standardization tests using analytical methods have identified three compounds A, B and C whose structure determination is in progress. These compounds could be used as markers for quality control of the prototype antihypertensive phytodrugs developed. Keywords: Anogeissus leiocarpa , Chemical Markers, Total Polyphenols, Total Flavonoids, Antioxidant Activity, HLC Profile pharmaceuticals or a combination of both for the treatment of 1. Introduction long-term diseases [3-5]. Hypertension is one of these The use of herbal medicines for primary care is central to diseases that is constantly increasing in many countries. In the global health system. In developing countries, it meets Burkina Faso, this disease has become a public health both economic and socio-cultural requirements. It is problem with a prevalence of 20% [6]. It represents the major estimated that more than 80% of the world's population now risk of cardiovascular disease and cerebrovascular accidents depends on medicinal plants for primary health care [1, 2]. In [7, 8]. Hypertension causes serious damage to vital organs addition, many people prefer phytotherapy to such as the brain, kidneys, heart, etc., and thus reduces life American Journal of Applied Chemistry 2020; 8(1): 6-12 7 expectancy. It is now widely accepted that highly reactive lyophilized extract was dissolved in 40 ml of distilled water. chemical species known as free radicals appear to be one of Using a separating funnel, successive runs were made with the major causes of this disease [9]. Since the treatment is solvents of increasing polarity (hexane, dichloromethane, long-term, in low-income countries most patients prefer ethyl acetate and butanol). herbal therapy. It is within this framework that pharmaceutical drugs are generally substituted by accessible 2.2.3. Phytochemical Screening recipes of traditional medicine whose effectiveness and Phytochemical screening was carried out on safety of use have been proven. The efficacy of plants in chromatoplaques (60 F254, glass support 5 x 10 cm, Fluka - treating hypertension is confirmed in many experimental Silica gel) according to the methods described in the studies [2, 3, 10]. Nowadays, several plant species are used in literature [19, 22, 23]. Each dry extract is solubilized in its the treatment of various diseases including hypertension. extraction solvent at concentration of 10 mg/mL and 5 µL is Thus, among the medicinal plants used in the treatment of deposited on the Thin Layer Chromatography plate. The hypertension by traditional health practitioners in Burkina different chromatograms have been developed in the Faso, we find Anogeissus leiocarpa . Anogeissus leiocarpa following solvent systems (Table 1). (Combretaceae) , is an evergreen tree widely distributed in Table 1. Solvent system used per extract. Africa [2, 11]. This plant is well known in traditional African medicine to treat many diseases, mainly skin diseases, Extracts Solvent system (V/V/V) hepatitis, hypertension, haemorrhoids, respiratory diseases, Hexane Hexane- ethyl-methanol acetate (7-2-1) Dichloromethane Toluene ethyl acetate acetic acid (5-4-1) headaches, etc. [12-16]. It has also shown strong antiradical Ethyl acetate Ethyl acetate - methanol - water (7-2-1) and antibacterial activity, antifungal activity against Butanol Ethyl acetate - methanol - water (7-2-1) pathogenic microorganisms [17-20]. Antiradical activity could therefore explain the use of this plant in the traditional 2.2.4. Total Phenolics Content treatment of hypertension. The general objective of this study Total phenolic content were determined by Singleton is to identify the fraction containing bioactive compounds, to method [24]. These compounds react with the Folin evaluate the content of phenolic compounds, in particular Ciocalteu reagent (FCR) in an alkaline medium. The loss of a flavonoids, as chemicals with antihypertensive potential and phenolic proton in an alkaline medium leads to a phenolate to identify compounds that can be isolated. The compounds anion that is capable to reducing. the FCR in molybdate identified will be studied in depth in order to elucidate their forming a blue colored molybdenum oxide complex with a chemical structure for the quality control and standardization maximum absorption at 760 nm. The intensity of the blue of the Phytomedicine prototype. coloration is proportional to the amount of total phenolics present in the test sample. The reaction mixture consisted of 2. Materials and Methods 1 ml extract, 1 ml 2N FCR and 3 ml 20% sodium carbonate solution. It was left at rest at room temperature for 40 min 2.1. Plant Material and then the absorbance is measured at 760 nm on the spectrophotometer (Agilent 8453). In the white control tube, The plant material consists of trunk bark of Anogeissus the extract was replaced by distilled water. A standard curve leiocarpa (Combretacaea). The harvested plant material was will be drawn with tannic acid (1-5 µg/ml). The tests were identified by the Laboratory of Ecology of the University carried out in triplicate. The total phenolic concentration of Joseph Ki-Zerbo. The collected bark was dried and then the extract was provided by the formula (1): powdered. This powder was used for this study. Tp T = (C tube x D)/ Ci (1) 2.2. Methods TPT is the total phenolic content of the extract expressed as 2.2.1. Moisture Content tannic acid equivalent (TEA)/g; CTube is the concentration in A powder sample Po (Po = 1 g) in a the crucible is dried in mg TEA/ml in the assay tube; D the dilution factor and Ci the an oven set at 105 °C for 3 hours. After cooling in a concentration in mg/ml in the stock solution. desiccator, weigh the dry test sample and note the mass P obtained. The drying process is repeated until the moisture 2.2.5. Total Flavonoids Content content attained a constant value [21]. The flavonoid assay was performed using the Kumaran method [25] adapted by Abdel-Hameed [26]. 2 ml of extract 2.2.2. Extraction Methods of concentration 1 mg/ml in methanol were mixed with 2 ml The decoction: 50 g of vegetable powder was dispersed in of 2% aluminium trichloride in methanol. After 40 minutes, 500 ml of distilled water. This was macerated for 15 minutes the absorbance was measured at 415 nm using the and then brought to the boil for 30 minutes. After cooling, spectrophotometer (Agilent 8453). The white control tube the mixture is filtered. The filtrate obtained was centrifuged holds 2 ml of methanol. A calibration curve has been drawn at 2000 rpm for 10 minutes. The supernatant obtained was using a series of standard solution of quercetin. The tests lyophilised and stored for later analysis. For the tests,